PIMEX-ARAUCO Program, Faculty of Natural Sciences and Oceanography, University of Concepción, Concepción, Chile.
Department of Zoology, Faculty of Natural Sciences and Oceanography, University of Concepción, Concepción, Chile.
Fish Physiol Biochem. 2010 Sep;36(3):757-765. doi: 10.1007/s10695-009-9350-6. Epub 2009 Aug 15.
We sought to provide a useful indicator of the presence of endocrine-disrupting contaminants along the marine coast of the South Pacific using Chilean flounder (Paralichthys adspersus). In light of the lack of information on vitellogenin for this species, we induced, purified, and identified the plasma vitellogenin of Chilean flounder inhabiting the Chilean coast. Vitellogenin (Vg) from Chilean flounder was purified by size exclusion and ion-exchange chromatography using plasma from juvenile males induced by injecting 17beta-estradiol. The Vg was detected by SDS-PAGE and Western blot analyses using an antibody against turbot (Scophthalmus maximus) vitellogenin. These analyses revealed a protein band of 205 kDa and three minor bands of 120, 90, and 68 kDa. These proteins were identified as Vg by means of mass spectrometry (LCQ Duo ESI-IT-MS), matching sequences of tryptic peptides to known sequences for several other fish species. The matches showed the presence of vitellogenin (VgI, VgII, Vg A and Vg B) in Chilean flounder, similar to species such as mummichog (Fundulus heteroclitus), Japanese medaka (Oryzias latipes), and white perch (Morone americana). These results are discussed in terms of identifying Vg in Paralichthys adspersus with the antibody to turbot Vg. Moreover, we compare the molecular size of Vg from Chilean flounder (large) with that of other flatfish species. Finally, we discuss the potential use of this molecule as a biomarker for the presence of xeno-estrogenic compounds along the Chilean coastline.
我们试图利用智利比目鱼（Paralichthys adspersus）为南太平洋沿海水域的内分泌干扰污染物提供一个有用的指标。鉴于该物种缺乏卵黄蛋白原的信息，我们诱导、纯化并鉴定了栖息在智利沿海的智利比目鱼的血浆卵黄蛋白原。通过用 17β-雌二醇注射诱导的幼鱼雄性血浆，使用大小排阻和离子交换层析从智利比目鱼中纯化卵黄蛋白原（Vg）。使用针对大菱鲆（Scophthalmus maximus）卵黄蛋白原的抗体通过 SDS-PAGE 和 Western blot 分析检测 Vg。这些分析显示出 205 kDa 的蛋白质带和三个较小的 120、90 和 68 kDa 的蛋白质带。这些蛋白质通过质谱法（LCQ Duo ESI-IT-MS）被鉴定为 Vg，与几种其他鱼类物种的胰蛋白酶肽的已知序列相匹配。这些匹配表明智利比目鱼中存在卵黄蛋白原（VgI、VgII、Vg A 和 Vg B），类似于美洲蟾鱼（Fundulus heteroclitus）、日本青鳉（Oryzias latipes）和白鲈（Morone americana）等物种。根据使用大菱鲆卵黄蛋白原抗体鉴定智利比目鱼中的 Vg 的结果，我们对这些结果进行了讨论。此外，我们比较了智利比目鱼（大）与其他比目鱼物种的 Vg 的分子大小。最后，我们讨论了将这种分子用作智利海岸线存在外源性雌激素化合物的生物标志物的潜力。
